Relationship between apoptosis and the BH2 domain sequence of the VP5 peptide of infectious pancreatic necrosis virus
Journal Title: Revista MVZ Cordoba - Year 2014, Vol 19, Issue 1
Abstract
Objective. To determine whether the level of apoptosis induced by infectious pancreatic necrosis virus (IPNV) is related to the amino acid sequence of the BH2 domain of the VP5 protein and the level of infectivity. Materials and methods. Three IPNV strains were used, the VP2 protein gene was amplified for genotyping and the VP5 sequence was also obtained. The infectivity of the strains was calculated using the viral titer obtained at 12, 24, 36 and 45 hpi in CHSE-214 cells. The percentage of apoptosis in infected cells was visualized by TUNEL assay and immunohistochemistry (caspase 3 detection). Results. The V70/06 and V33/98 strains corresponded to genotype Sp, while V112/06 to VR-299; the amino acid analysis of the V70/06 strain allows its classification as middle virulent strain and V33/98 and V112/06 strains as low virulent ones; infection with the V112/06 strain produced a lower viral titer (p<0.05). The VP5 gene of the 3 strains showed four homologous domains to Bcl-2, however, the BH2 domain was truncated in V70/06 and V33/98 (12 kDa), being complete (15kDa) in V112/06, which also showed the Trp155 residue, equivalent to Trp188 considered as a critical factor for the function of Bcl-2. The average apoptosis was below 12%, showing no differences between strains (p>0.05). Conclusions. The results showed that the differences in the BH2 sequence of the VP5 protein, infectivity and the VP2 sequence are not associated with the modulation of apoptosis.
Authors and Affiliations
Cesar Ortega S, Ph. D *| Universidad Autonoma del Estado de México, Faculty of Veterinary Medicine and Animal Husbandry, Advanced Animal Health Research and Study Center. Toluca, Mexico, AP. 4-56.Correspondence: cos_mx@hotmail.com, Sylvia Rodríguez S, Ph. D| Biologic Research Center (High Council for Scientific Research) c/ Ramiro de Maeztu 9 Madrid 28040, Spain., Juan Carlos Espinoza, M. Sc| Universidad de Valparaiso, Faculty of Sciences, Biochemistry and Virology Laboratory. Gran Bretaña 1111, Valparaíso, Chile., Juan Kuznar, Ph. D| Universidad de Valparaiso, Faculty of Sciences, Biochemistry and Virology Laboratory. Gran Bretaña 1111, Valparaíso, Chile., Alex Romero, Ph. D| Universidad Austral de Chile, Faculty of Veterinary Sciences, Biotechnology and Aquatic Pathology Laboratory. Valdivia, Chile.Centro FONDAP: Interdisciplinary Center for Aquaculture Research (INCAR), Chile., Ricardo Enríquez, Ph. D| Universidad Austral de Chile, Faculty of Veterinary Sciences, Biotechnology and Aquatic Pathology Laboratory. Valdivia, Chile.
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